Release Kinetics of Nisin from Chitosan–Alginate Complex Films

Understanding the release kinetics of antimicrobials from polymer films is important in the design of effective antimicrobial packaging films. The release kinetics of nisin (30 mg/film) from chitosan–alginate polyelectric complex films prepared using various fractions of alginate (33%, 50%, and 66%) was investigated into an aqueous release medium. Films containing higher alginate fractions showed significantly lower (P < 0.05) degree of swelling in water. Total amount of nisin released from films into an aqueous system decreased significantly (P < 0.05) with an increase in alginate concentration. The mechanism of diffusion of nisin from all films was found to be Fickian, and diffusion coefficients varied from 0.872 × 10^?9 to 8.034 ×10^?9 cm²/s. Strong complexation was confirmed between chitosan and alginate polymers within the films using isothermal titration calorimetry and viscosity studies, which affects swelling of films and subsequent nisin release. Complexation was also confirmed between nisin and alginate, which limited the amount of free nisin available for diffusion from films. These low‐swelling biopolymer complexes have potential to be used as antimicrobial packaging films with sustained nisin release characteristics.     

Alginate–calcium coating incorporating nisin and EDTA maintains the quality of fresh northern snakehead (Channa argus) fillets stored at 4 °C

BACKGROUND: Antimicrobial packaging is a novel food packaging technology for controlling the growth of food‐borne pathogens or spoilage bacteria in ready‐to‐eat food products. Fresh fish are highly perishable foodstuffs and are extremely susceptible to microbial activities. An alginate–calcium coating incorporating nisin and ethylenediaminetetraacetic acid (EDTA) was used as an antimicrobial packaging to maintain the quality of northern snakehead (Channa argus) at refrigeration temperature (4 ± 1 °C). Northern snakehead fillets were left untreated (CK), or were treated with 1000 IU mL^?1 nisin and 150 µg mL^?1 EDTA (T1), alginate–calcium coating (T2), or alginate–calcium coating incorporating 1000 IU mL^?1 nisin and 150 µg mL^?1 EDTA (T3). RESULTS: All treatments retarded the decay of the fish fillets. T1 more efficiently inhibited the growth of total viable mesophilic bacteria (P < 0.05) and total psychrophilic bacteria (P > 0.05) than did T2 or T3. Coating treatments predominantly reduced chemical spoilage, reflected in total volatile base nitrogen (P > 0.05), trimethylamine (P > 0.05), pH (P < 0.05), and thiobarbituric acid (P < 0.05), retarded water loss (P < 0.05), and increased the overall sensory scores of fish fillets (P < 0.05) compared with those of T1. There was no significant difference between the coating treatments T2 and T3 (P > 0.05). CONCLUSION: Alginate–calcium coating treatments efficiently enhanced the quality of northern snakehead fillets during storage. Copyright © 2009 Society of Chemical Industry     

Application of chitosan–alginate microspheres for the sustained release of bacteriophage in simulated gastrointestinal conditions

This study was designed to evaluate the acid stability, release property and antimicrobial efficacy of Escherichia coli O157:H7 bacteriophages encapsulated in chitosan–alginate microspheres under the simulated gastrointestinal conditions. The bacteriophages belonging to Myoviridae family were stable at the pH above 4 in trypticase soy broth. The chitosan–alginate microspheres exhibited protective effect on the viability of bacteriophages in the simulated gastric conditions at pH 2.0 and pH 2.5, showing 4.8 and 5.6 log PFU mL^‐1, respectively, after 1 h of incubation at 37 °C. The release per cent of bacteriophages from microspheres gradually increased up to 65% in the simulated intestinal condition (pH 7.5) at 37 °C for 6 h. The lytic efficacy of chitosan‐ and alginate‐encapsulated bacteriophages against E. coli O157:H7 was significantly maintained in the simulated intestinal conditions to 10 h of incubation (1.3 log reduction). The results suggest that the chitosan–alginate microspheres can be used as a reliable delivery system for bacteriophages.     

Reduction of Listeria monocytogenes in cold‐smoked salmon by bacteriophage P100, nisin and lauric arginate,singly or in combinations

Three GRAS antimicrobials including, lauric arginate (LAE), bacteriophage P100 (phage P100) and bacteriocin nisin, were evaluated either singly or in combinations for the reduction of initial load of Listeria monocytogenes in cold‐smoked salmon (CSS). The stability of phage P100 in the presence of LAE (200 ppm) and nisin (500 ppm) or at 10× and 100× of these concentrations was determined at 4 °C or 30 °C for 24 h in a broth model. Phage P100 was found to be highly stable in the presence of these antimicrobial agents as plaque‐forming units (PFU) did not vary between control and antimicrobial‐treated phage. The survival of L. monocytogenes in the presence of phage P100, nisin and LAE showed remarkable reduction within 24 h both at 4 °C or 30 °C in broth. Treatment of CSS containing 3.5 log CFU cm^?2 L. monocytogenes with phage P100 (10⁸ PFU mL^?1), nisin (500 ppm) and LAE (200 ppm) showed strong listericidal action and reduced the L. monocytogenes by 2–3 log CFU cm^?2 after 24 h. Among the combined treatments, phage P100 + LAE or nisin + LAE exhibited the most listericidal action in which L. monocytogenes cells were reduced to undetectable level within 24 h in CSS.     

Structure evolution of pullulan–alginate edible films during drying studied by low‐field NMR

The evolutions of spatial structure for pullulan, alginate, and pullulan–alginate blend aqueous solutions during drying were investigated by low‐field nuclear magnetic resonance. For pullulan solutions, during the first 1,320 min of drying, NMR signal intensity in profiles did not vary with position. Thereafter, the apparent shift of the air–pullulan interface began at 1,440 min, indicating the onset of progressive shrinkage of pullulan samples. Similar shrinkage phenomena were observed for alginate and pullulan–alginate blend solutions. In contrast, spatial structural heterogeneity was observed for alginate solution during drying time from 1,200 to 1,560 min, due to the formation of a skin layer near the air–alginate interface. Based on the change of polymer solutions during drying, two‐stage evaporation process was detected in the aqueous pullulan, alginate, and their blend solutions. Moreover, the evaporation rate of water during the second stage of drying was significantly lower than that of the first stage.

Practical applications

Edible films have emerged as an alternative to synthetic petroleum‐based polymers for food packaging. The evolutions of spatial structure of pullulan–alginate solutions during drying were studied by monitoring of their NMR profiles. Information shown in this study would provide some scientific basis for studying film‐forming mechanism of edible films and their applications in the food field.     

Encapsulationof lycopene from watermelon in calcium‐alginate microparticles using an optimised inverse‐gelation method by response surface methodology

Lycopene exhibits strong antioxidant activity due to its unsaturated molecular bonds, which also contributes to its susceptibility for degradation. Encapsulation techniques can reduce lycopene degradation, increasing its potential applications in functional foods and nutraceuticals. The objective of this study was to optimise the encapsulation of lycopene from watermelon in alginate microparticles using the inverse gelation method. Box–Behnken design was used for the optimisation of three variables: concentrations of alginate (w/v %) and CaCl₂ (g L^?1), and gelation time (min). Two types of alginate were investigated (low viscosity and high viscosity) and optimised separately using encapsulation efficiency and loading capacity as responses. Results indicated that the models had a good fit to the experimental data and the optimal conditions varied depending on the type of alginate. In general, particles prepared with low‐viscosity alginate exhibited higher encapsulation efficiency and loading capacity and could be used for further research.     

The rheological property,antimicrobial stability and release evaluation of soya protein isolate/alginate‐based films incorporated with thymol

Soya protein isolate (SPI)/alginate (ALG)/montmorillonite (MMT)/thymol (THY) nanocomposite films were prepared with a codrying process to enhance the antimicrobial stability. The results showed higher gelation point temperature and more stable gelling network were obtained by the addition of MMT. Better thermal stability and sustained release effect was found in the coblended film containing MMT, in which THY release rate at 25 °C decreased with increasing MMT concentration but increased with temperature at a constant relative humidity. The SPI/ALG/THY, SPI/ALG/THY‐CO and SPI/ALG/THY‐CO‐0.6 (MMT content = 0.6 g g^?1 SPI) films effectively inhibited the growth of Escherichia coli by 0.95, 0.91 and 0.70 log cfu mL^?1 and of Staphylococcus aureus by 0.94, 0.89 and 0.73 log cfu mL^?1, respectively, on the surface of raw sausage, compared to the control film.     

Preparation and optimization of soy protein isolate–high methoxy pectin microcapsules loaded with Lactobacillus delbrueckii

This study was aimed to use soy protein isolate (SPI) and high methoxy pectin (HMP) as encapsulating materials for probiotic bacterial (Lactobacillus delbrueckii) delivery systems. The encapsulation conditions were optimised, and scanning electron microscopy (SEM) was used to characterise the microstructural changes of the microcapsule. The results showed that the optimal conditions for microcapsule preparation were 90 mg mL^?1 SPI and 1 mg mL^?1 HMP, with a SPI/HMP ratio of 7:1 (v/v), and a L. delbrueckii suspension to SPI–HMP complex ratio of 1:1 (v/v). The viability of the probiotics in the microcapsules reaching the small intestine was 3 log CFU mL^?1 higher than that of naked bacteria. SEM showed that the surface of the SPI–HMP compound microcapsules was smooth and that a large number of L. delbrueckii could be seen in cross‐sections of the microcapsules.     

Efficacy of Acorus calamus L. essential oil as a safe plant‐based antioxidant,Aflatoxin B1 suppressor and broad spectrum antimicrobial against food‐infesting fungi

The study explores the efficacy of Acorus calamus L. essential oil (EO) as a safe plant‐based broad spectrum antifungal, antiaflatoxin, antioxidant food additive. The oil completely inhibited the growth and toxin production of the toxigenic strain of Aspergillus flavus at 0.4 and 0.25 μL mL^?1, respectively. EO exhibited pronounced antifungal activity against sixteen food‐infesting fungal species at 0.5 μL mL^?1. The EO showed strong antioxidant efficacy (IC₅₀ 1.06 μL mL^?1) and nonphytotoxic nature on germination of chickpea seeds. The EO was found nonmammalian toxic showing high LD₅₀ (4877.4 μL kg^?1) for mice (oral, acute). The chemical profile of EO was determined through GC and GC–MS analysis. The findings strengthen the possibility of A. calamus EO as a plant‐based food additive in view of its favourable safety profile, antioxidant and antiaflatoxigenic efficacy and broad spectrum antimicrobial activity against food‐infesting fungi.     

An alternative encapsulation approach for production of active chitosan–propolis beads

Encapsulation is a promising technology to carry natural active substances, preventing their loss and maintaining their stability until use. Beads of chitosan‐containing propolis have been prepared using a mono‐pore filter device, which permits the encapsulation of natural polyphenols avoiding heat treatments, high shear rates and the use of toxic solvents. Beads proved to be active against Bacillis cereus, Escherichia coli, Listeria innocua, Pseudomonas fluorescens, Yarrovia lipolytica and three moulds strains; the highest effect was found against Staphylococcus aureus (MIC 0.8 mg beads mL^?1). Results in liquid cultures of S. aureus evidenced that beads were able to release the flavonoids from propolis: the diffusion of the active compounds is a key factor in the exploitation of the microbial activity. The obtained chitosan–propolis beads represent an example of natural antimicrobial delivery system that could be used to prevent the growth of pathogenic/spoilage bacteria in food applications.     

Pea protein‐based capsules for probiotic and prebiotic delivery

Capsule design for entrapping Bifidobacterium adolescentis and fructoligosacchardes within a pea protein isolate–alginate matrix was investigated as a function of total biopolymer concentration (2.5–6.5%), fructoligosacchardes levels (0–3.0%) and needle gauge (G; 20 vs. 27) by extrusion. Capsules were accessed based on the levels of entrapped fructoligosacchardes and protein, size, swelling and probiotic survival within simulated gastric juice (SGJ) and release properties within simulated intestinal fluids (SIF). All designs offered sufficient protection to B. adolescentis during challenge experiments and showed evidence of burst and prolonged release within SIF. Capsule size was found to depend on needle gauge, with diameter of 2.17 and 1.75 mm for those produced by the 20 and 27 G needles, respectively. All designs were found to shrink once immersed in SGJ. Encapsulation of probiotics using plant‐based materials could play a role in food applications and as a feed/pet food supplement.     

Antibiofilm formation and anti‐adhesive (to HEp‐2 cells) effects of rosemary water extract against some food‐related pathogens

The present work aimed to determine the bioactive compounds in two rosemary water extracts (RWE1 and RWE2) and to assess their antimicrobial, anti‐adhesive and antibiofilm potentials against the food‐related Bacillus and Pseudomonas species at concentrations; 4, 8, 12, 16 and 20 mg mL^?1. Phenolic compounds and isoflavones in the RWEs were determined using HPLC. The concentrations of most bioactive compounds of RWE1 (benzoic, ellagic, gallic and rosmarinic acids, daidzein and genistein) were higher than that of RWE2. The MIC₉₀ of RWE1 and RWE2 against all tested bacteria was 12 and 16 mg mL^?1, respectively. The anti‐adhesive and antibiofilm doses were higher than MIC₉₀. RWE1 and RWE2 showed potential reduction in the bacterial cell adhesion to HEp‐2 cells – 17.5–64.7 and 12.2–52.9%, respectively. In conclusion, this study emphasises the effective use of RWE as a natural anti‐adhesive and antibiofilm agent against Bacillus and Pseudomonas, without difficult extraction procedure.     

Enhanced antimicrobial activity of nisin in combination with allyl isothiocyanate against Listeria monocytogenes,Staphylococcus aureus,Salmonella Typhimurium and Shigella boydii

This study was designed to evaluate the synergistic antimicrobial effect of nisin and allyl isothiocyanate (AITC) against Listeria monocytogenes, Staphylococcus aureus, Salmonella Typhimurium and Shigella boydii. The synergistic interactions between nisin and AITC were observed against all foodborne pathogens, showing the fractional inhibitory concentrations <1. The populations of L. monocytogenes and S. aureus at the combined treatment of nisin and AITC were decreased to below 1 log CFU mL^?1 after 10‐h incubation at 37 °C. The changes in fatty acid profiles of all strains were substantially influenced by nisin alone and the combined treatment of nisin and AITC. A good agreement was observed among cell viability, membrane permeability and depolarisation activity in response to nisin and AITC. The results suggest that nisin and AITC as synergistic inhibitors could be an effective approach to achieve satisfactory antimicrobial activity against a wide range of foodborne pathogens.     

Hurdle Effect of Antimicrobial Activity Achieved by Time Differential Releasing of Nisin and Chitosan Hydrolysates from Bacterial Cellulose

We investigated the combined antimicrobial effect of nisin and chitosan hydrolysates (CHs) by regulating the antimicrobial reaction order of substances due to differential releasing rate from hydroxypropylmethylcellulose‐modified bacterial cellulose (HBC). The minimum inhibitory concentration of nisin against Staphylococcus aureus and that of CHs against Escherichia coli were 6 IU and 200 μg/mL, respectively. Hurdle and additive effects in antimicrobial tests were observed when nisin was used 6 h before CH treatment against S. aureus; similar effects were observed when CH was used before nisin treatment against E. coli. Simultaneously combined treatment of nisin and CHs exhibited the low antimicrobial effect. HBC was then selected as the carrier for the controlled release of nisin and CHs. A 90% inhibition in the growth of S. aureus and E. coli was achieved when 30 IU‐nisin‐containing HBC and 62.5 μg/mL‐CH‐containing HBC were used simultaneously. The controlled release of nisin and CHs by using HBC minimized the interaction between nisin and CHs as well as increased the number of microbial targets.     

Growth patterns of Escherichia coli O157:H7 in ground beef treated with nisin,chelators, organic acids and their combinations immobilized in calcium alginate gels

The effects of antimicrobial substances including nisin, acetic acid, lactic acid, potassium sorbate and chelators (disodium ethylenediamine tetraacetic acid [EDTA] and sodium hexametaphosphate [HMP]), alone or in combination and, with or without immobilization in calcium alginate gels, on the growth of Escherichia coli O157:H7 in ground beef were investigated. Results showed that acetic acid and potassium sorbate could inhibit the growth of E. coli O157:H7 effectively at 10°C and at 30°C. Both EDTA and HMP did not halt the growth of E. coli O157:H7. In an antimicrobial system immobilized with calcium alginate, most of the antimicrobials could not inhibit the growth of E. coli O157:H7 in ground beef at 10°C and at 30°C, with the exception of acetic acid and lactic acid. Immobilization did not enhance the effectiveness of acetic acid against E. coli O157:H7 in ground beef at 10°C and at 30°C (P>0.05) but it did enhance the effectiveness of lactic acid at 10°C. In a system combining different antimicrobials, treatment with nisin /EDTA or nisin/potassium sorbate at 10°C revealed a significantly lower population change of E. coli O157:H7 compared to samples treated with nisin, EDTA or potassium sorbate alone. The use of calcium alginate immobilization further enhanced the effectiveness of the combination system of nisin/EDTA, nisin/acetic acid and nisin/potassium sorbate on the growth of E. coli O157:H7 in ground beef at 10°C but it was not effective at 30°C.     

Effects of simulated gastrointestinal digestion on the physicochemical properties,erythrocyte haemolysis inhibitory ability and chemical antioxidant activity of mulberry leaf protein and its hydrolysates

Proteins and peptides must be degraded and modified in the gastrointestinal (GI) tract prior to absorption; this process changes their physicochemical and biological properties. Mulberry leaf protein (MP) and its hydrolysates (HMP) have favourable antioxidant activities. To investigate, in vitro and separately, the effects of GI digestion and intestinal digestion on the stability of MP and HMP, we monitored the changes in secondary structures, amino acids, molecular weights and antioxidant activities. We found that MP was more hydrolysed by pepsin than by pancreatin, unlike HMP. The final digests of MP and HMP were mainly composed of polypeptides (0.5–6.5 kDa) and oligopeptides (<0.5 kDa), respectively. The GI digestion influenced MP and HMP differently; GI digestion increased the antioxidant efficiency of MP and decreased that of HMP. For the intestinal digests, the antioxidant activities of MP and HMP also differed. The 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) and 2,2′‐azinobis (3‐ethylbenzothiazoline‐6‐sulphonic acid) diammonium salt (ABTS) radical quenching abilities of MP and HMP at 1 mg mL^?1 were comparable to or exceeded that of L‐glutathione (GSH) and 6‐hydroxy‐2,5,7,8‐tetramethychroman‐2‐carboxylic acid (trolox). Meanwhile, the erythrocyte haemolysis rates of MP, HMP and their GI products at 0.05–1.0 mg mL^?1 were significantly lower than that of the 2,2′‐azobis (isobutyramidine) dihydrochloride (AAPH) control. Both MP and HMP can be used as natural antioxidants and may promote digestive health.     

Release Kinetic in Yogurt from Gallic Acid Microparticles with Chemically Modified Inulin

Gallic acid (GA) was encapsulated with native (NIn), cross‐linked (CIn) and acetylated (AIn) inulin by spray‐drying. Inulin microparticles were characterized by encapsulation efficiency (EE) and their release profile in yogurt. The EE was significantly higher for GA‐CIn (98%) compared with GA‐NIn (81%) and GA‐AIn (77%) microparticles, showing the effect of the modification of inulin on interaction of GA‐polymer. GA release profile data in yogurt for GA‐CIn, GA‐NIn and GA‐AIn were fitted to Peppas and Higuchi models in order to obtain the GA release rate constant. Although the GA release rate constants were significantly different among systems, these differences were slight and the GA release was fast (80% < 2 h) in the three systems, showing that inulin‐systems did not control GA release in yogurt. The mechanism of GA release followed a Fickian diffusion and relaxation of chains for all microparticles. According to the release profile, these microparticles would be best suited for use in instant foods.     

Ultrasound‐assisted extraction of polyphenols from potato peels: profiling and kinetic modelling

Ultrasound‐assisted extraction (UAE) at 33 and 42 kHz has been investigated in the extraction of polyphenols from peels of two potato varieties, cream‐skinned Lady Claire (LC) and pink‐skinned Lady Rosetta (LR), commonly used in snack food production. Extraction efficacy between the UAE‐untreated (control) and the UAE‐treated extracts was assessed on the total phenolic content and antioxidant capacities (DPPH and FRAP). Application of UAE showed significantly higher recovery of phenolic compounds compared to solid–liquid extraction process alone. Lower ultrasonic frequency (33 kHz) was more effective in recovering polyphenols compared to 42 kHz ultrasonic treatment. The liquid chromatography‐tandem mass spectrometry revealed that chlorogenic acid and caffeic acid were the most prevalent phenolics in LR peels, whereas caffeic acid was dominant in LC peels. Peleg's equation showed a good correlation (R² > 0.92) between the experimental values and the predicted values on the kinetics of UAE of phenolic compounds.     

Release and antilisterial properties of nisin from zein capsules spray-dried at different temperatures

Nisin is an effective antimicrobial against a broad spectrum of Gram-positive bacteria. It has been proposed that reduced efficacy of nisin in foods can be improved by technologies such as encapsulation to protect it from interferences by food matrix components. The potential of spray drying, a practical technology, was studied in this work for encapsulation of nisin in zein microcapsules at four inlet temperatures between 75 and 120 °C. At 95 °C and above, no apparent loss of nisin activity was observed after spray drying. At pH 6.0, burst release of nisin was impacted by spray drying temperature more than equilibrium release, possibly due to influences on capsule structures. At pH 2.0, complete release of nisin in 30 min was observed, contrasting to limited release over 8 d at pH 8.0. Capsules produced at an inlet temperature of 105 °C showed the most sustained release of nisin at pH 6.0. For these capsules, sustained release of nisin to >80% was observed at pH 6.0 and 8.0 when NaCl was used at 0.5 mol/L. Finally, at 400 IU/mL, the encapsulated nisin demonstrated slightly improved antilisterial properties than free nisin in reduced fat (2g/100g) milk but much work is still needed to enhance the antimicrobial effectiveness.     

Preparation and optimisation of liposome‐in‐alginate beads containing oyster hydrolysate for sustained release

Oyster (Crassostrea gigas) hydrolysate shows antihypertensive effect in our previous study. Oral administration of oyster hydrolysate can loss bioactive peptides due to enzymatic degradation in vivo. To maximise its bioavailability, liposome‐in‐alginate (LA) beads were used to encapsulate the oyster hydrolysates to protect from degradation and obtain sustained release. The preparation conditions of the LA beads were optimised by response surface method using a model peptide of tyrosylalanine (YA). Their characterisation, swelling and release properties were investigated. The optimised conditions for the concentration of calcium chloride, sodium alginate and the amount of ethanol‐dissolved lecithin (EDL) were 0.5 m , 3% and 95.4 mg, respectively. The encapsulation efficiencies of YA and the oyster hydrolysate in the optimised condition were 74.9% and 84.3%, respectively. The release time of the oyster hydrolysate in the simulated gastrointestinal fluid was up to 16 h. The LA beads can be recommended to encapsulate oyster hydrolysate for bioavailability improvement.